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Biomedical subjects

C Campanella

Publications and source records attributed to C Campanella.

At least 37 records · Page 2Linked to original sources

A novel protein cross-reacting with antibodies against spectrin is localised in the nucleoli of amphibian oocytes.

Cytoskeletal proteins such as actin and myosin are important constituents of the nucleoplasm. Spectrin is an actin binding protein typically related to plasma membrane; recently, it has been found that it is widespread and forms distinct membrane protein domains in such organelles as the Golgi. In this paper, the large germinal vesicle of amphibian oocytes was chosen as a particularly suitable system to investigate the presence and location of spectrin in the nucleus. We manually isolated the germinal vesicles of both Discoglossus pictus and Xenopus laevis oocytes, and processed them for SDS-PAGE, immunoblotting and immunoprecipitation. By the use of an antibody against the general form of brain beta spectrin (betaIIsigma1) and of an anti-alpha brain spectrin (alphaIIsigma*), a band of 230 kDa was identified as a nuclear spectrin-like molecule. Moreover the 230 kDa protein was extracted from the nuclei by 1 M KCl, similarly to spectrin in other systems. In oocyte sections and nuclear spreads incubated with anti-alphaIIsigma* and/or anti-betaIIsigma1 antibodies, the immunostain was localised in the nucleoplasm and in the outer shell of the round bodies abundantly present in the germinal vesicle. Sections of the same oocytes, stained with a monoclonal antibody against nucleolar fibrillarin and anti-alphaIIsigma*, showed co-localisation of the two antibodies. It was concluded that, in the germinal vesicle of amphibian oocytes, a spectrin-like molecule is a part of the outer shell of nucleoli. It is hypothesised that spectrin, together with actin, might be instrumental in keeping nucleoli attached to the inner nuclear membrane, as nucleoli migrate during oogenesis to the inner aspect of the nuclear envelope, where they are stably kept until the end of their growth. Furthermore, these results strongly suggest that the 230 kDa band might comprise both an alpha and a beta chain of the same apparent molecular mass, thus constituting a novel form of a spectrin-like molecule.

Animals↗

Latissimus dorsi muscle: assessment of stimulation after cardiomyoplasty with Doppler US tissue imaging.

PURPOSE: To evaluate Doppler ultrasound (US) tissue imaging for assessment of stimulated skeletal muscle contraction. MATERIALS AND METHODS: Seven patients were studied after left latissimus dorsi cardiomyoplasty. Myograft contraction programmed on alternate cardiac cycles was assessed with Doppler US tissue imaging. Beat-to-beat variation in inferior wall motion was assessed by examining peak myograft velocities during 10 muscle-assisted and 10 nonassisted cardiac cycles. The temporal relationship between electrostimulation and myograft contraction, changes in cardiac geometry, and the effect of alterations in stimulation voltage and muscle synchronization were assessed. RESULTS: Significant beat-to-beat variation in velocity profile could be detected in the proximal myograft in six patients (P < .05). Potentiation of infero-posterior wall motion was measurable in five patients (mean peak systolic wall velocity: nonassisted, 2.5 cm.sec-1 +/- 0.5 [standard deviation]; assisted, 7.8 cm.sec-1 +/- 6.3). The response between stimulation voltage and inferoposterior wall velocity was sigmoid. CONCLUSION: Doppler US tissue imaging depicted the effects of myostimulator programming on muscle contraction and ventricular wall motion.

Cardiomyoplasty↗

Optimizing muscle synchronization after dynamic cardiomyoplasty. Two educational cases.

Muscle stimulator programming is fundamental in ensuring optimal latissimus dorsi graft function after cardiomyoplasty. The ventricle-muscle synchronization delay has a significant effect on graft function, but can be difficult to optimize. The established method for ensuring appropriate synchronization involves the use of M-mode echocardiography. We present two educational cases in which M-mode and Doppler echocardiography have been instrumental in illustrating electrophysiologic and haemodynamic factors which need to be considered in evaluating an appropriate synchronization delay.

Atrial Fibrillation↗

Hufnagel revisited: a descending thoracic aortic valve to treat prosthetic valve insufficiency.

In 1953 Hufnagel and Harvey reported their successful treatment of aortic valve insufficiency by the implantation of a ball-valve prosthesis into the descending thoracic aorta. Since then, great advances in technology, surgery, and anesthesia have made aortic valve replacement a more common procedure with relatively low mortality. This remains true for the vast majority of prosthetic valve replacements. However, cases requiring reoperation can be difficult, leading to a much higher degree of morbidity and mortality. In selected patients who require repeated approaches to the aortic root we propose that Hufnagel's original idea may still be of value to reduce the severity of aortic insufficiency. We report our experience in 4 cases of aortic prosthetic incompetence, all of which were improved by two New York Heart Association functional classes after a modification of Hufnagel's procedure.

Adult↗

Retention of bioprosthetic valve annulus in mitral prosthetic replacement.

Mitral valve prosthetic replacement carries high mortality rates by modern standards, and mitral bioprostheses are particularly prone to degeneration. Bioprosthetic replacement may be technically difficult when there is calcification of the tissue ingrowth, strut incorporation, or valve-to-annulus size mismatch at the primary operation. A "valve-in-valve" technique is described where the mechanical prosthesis is implanted in the bioprosthetic annulus in order to avoid such difficulties. The results in the first eight patients are presented, showing post-operative Doppler-derived transvalvar mean gradients between 3.9 mmHg and 7.5 mmHg, and estimated valve areas between 1.9 cm2 and 3.5 cm2. All patients are alive at between 20 and 30 months (mean 23.6 months) after operation, they are without serious post-operative morbidity and are in functional class I or II (NYHA classification).

Aged↗

Jelly plug dissolution in Discoglossus pictus eggs (Anura) involves peroxidase-like activity and oxidative opening of disulphide bonds.

In amphibian eggs the formation of a capsular chamber is one of the most striking events occurring either upon oviposition or after fertilisation. In the egg of the anuran Discoglossus pictus a capsular chamber forms following fertilisation or activation; the egg with its vitelline envelope rotates in this chamber according to gravity. Previous work showed that the chamber is the product of plug dissolution. The plug is a lens-shaped jelly coat, typical of Discoglossus, covering only part of the animal hemisphere. Its dissolution is caused by material released from the egg about 15 min after fertilisation through exocytosis of at least two types of vacuoles. Liquefaction of the plug correlates with the reduction of disulphide bonds present in the jelly matrix. In this study we investigated the nature of the substances released from the egg and some changes occurring in the plug during liquefaction. SDS-PAGE showed that the proteic profile of the plug changes dramatically after fertilisation, confirming proteic cleavage in the plug matrix during its dissolution. Through in vitro tests and electrophoretic analysis of the Ringer solution in which the egg exudate was collected, an increase in the activity of the solution was determined in the presence of hydrogen peroxide, and peroxidase activity was depicted in the egg exudate. The presence of free thiol groups and cysteic acid residues (or cysteine sulphinic acid) in the plugs of activated eggs was established, suggesting that during plug dissolution some disulphide bonds are oxidatively opened. This suggests that enzyme(s) with peroxidase activity are released following fertilisation. We surmise that such enzymes are contained in the intraovular vacuoles the exocytosis of which triggers the onset of plug liquefaction. The possible release of hydrogen peroxide from the egg is discussed.

Amino Acids↗

Post-fertilization changes in Discoglossus pictus (Anura) eggs result in the formation of a capsular chamber where the egg rotates.

Discoglossus pictus is one of the few anurans with an egg where a capsular chamber forms as a consequence of fertilization; the egg with its vitelline envelope rotates in this chamber according to gravity. We investigated the formation of the capsular chamber through various experimental cytochemical and ultrastructural approaches, and found that it is the product of plug liquefaction. The plug is a lens-shaped jelly coat typical of Discoglossus, and covering only part of the egg animal half. About 15 min after fertilization, granular material coming from the egg enters the plug, which gradually dissolves and, once liquefied, reorganizes itself around the entire egg, thus forming the chamber. This process goes through stages of rearrangement of the 25-A- and 250-A-thick filaments which constitute the plug matrix. The material entering the plug derives from the exocytosis of two vacuole types, with electron transparent and granular PAS-positive contents. Liquefaction of the plug correlates with the reduction of disulfide bonds present in its matrix. Furthermore, in vitro tests showed that the substances released from the egg are active in selectively dissolving only the plug, and lose activity upon boiling.

Animals↗

Preliminary results of left heart bypass in pigs using a heparin-coated centrifugal pump.

To assess the feasibility of left ventricular assist without systemic heparinization, we used a commercially available (Sarns 3M) centrifugal pump with tubing set and cannulas, all internally precoated for the purpose of this study with heparin, to bypass the left ventricle in 12 pigs for periods of either 1 or 3 hours. There was no significant activation of clotting and there was no sign of generalized embolization. However, on postmortem studies, 5 kidneys out of 22 examined showed signs of minimal thromboembolism. This experiment shows that artificial left ventricular assist, free of systemic heparinization but using heparin precoating, is feasible and safe, at least for a short period of time.

Animals↗

Influence of obesity on the early and long term results of surgery for coronary artery disease.

In order to determine the effect of obesity on the results of coronary artery bypass graft (CABG) surgery, we compared 250 obese patients undergoing CABG procedures between 1984 and 1987 with 250 age- and sex-matched controls of normal body mass index (BMI) undergoing CABG in the same period. The obese group had a greater incidence of diabetes mellitus (p less than 0.02), hypertension (p less than 0.05), hyperlipidaemia (p less than 0.05), and left main stem coronary artery disease (p less than 0.001). No differences were identified in the surgery performed, but obesity was associated with prolonged total bypass time (p less than 0.05). Operative mortality was 0.8% in both groups. Multivariate analysis demonstrated obesity to be an independent risk factor for perioperative morbidity (p less than 0.05). Univariate: respiratory (p less than 0.01); leg wound (p less than 0.001); myocardial infarction (p less than 0.02); arrhythmias (p less than 0.02); sternal dehiscence (p less than 0.02). At a mean follow-up time of 36.9 months obese patients exhibited a greater incidence of significant recurrent angina (p less than 0.01), which was associated with further weight gain (mean 12.2 kg; linear correlation: p less than 0.001, r = 0.891). Although in CABG surgery operative mortality is not increased in obese patients, aggressive pre- and postoperative weight control is indicated to reduce both perioperative morbidity and the incidence of recurrent angina.

Angina Pectoris↗

[The centrifugal pump--a new method of assisting the failing heart following open heart surgery].

The centrifugal pump is a new mechanical ventricular assistant device, which is used to help the failing heart after cardiopulmonary bypass or postoperatively in the ICU. The centrifugal pump is hemodynamically able to improve the coronary and peripheral perfusion in patients in cardiogenic shock. The pump can displace the entire cardiac output and let the blood bypass the left ventricle, which gives the failing heart a possibility for metabolic correction and regeneration. Ten patients were included in the early personal experience at the cardio-thoracic unit in Edinburgh, with a survival rate of 50%, which is regarded as being very satisfactory. All hemodynamic parameters were normalised in the surviving patients.

Aged↗

Different cytoskeletal organization in two maturation stages of Discoglossus pictus (Anura) oocytes: thickness and stability of actin microfilaments and tropomyosin immunolocalization.

In Discoglossus pictus oocytes, the germinative area (GA) contains long and irregular microvilli where actin microfilaments are located. In the egg, the funnel-shaped dimple that originates by invagination of the GA is present. In the dimple both microvilli and microfilament bundles have a very orderly appearance. This report extends previous observations (Campanella and Gabbiani, Gamete Res 3:99-114, 1980) and shows that GA microfilaments are thinner (36 A average) than dimple microfilaments (60 A average), as measured in ultrathin section. Moreover, the interfilament distance is smaller in GA bundles than in the dimple bundles. To get an insight into actin organization in oocytes and eggs, we used an actin-depolymerizing factor (ADF) in which cryostat sections were incubated prior to immunofluorescent staining with antiactin antibodies. The microfilaments of the GA microvilli and partially of the oocyte cortex are resistant to ADF when compared to those in the dimple and the rest of the egg cortex. We also investigated immunocytochemically the presence of tropomyosin and found that this protein is localized in the dimple and in the cortex of oocytes and eggs but is absent in the GA.

Actin Cytoskeleton↗

Antispectrin antibodies stain the oocyte nucleus and the site of fertilization channels in the egg of Discoglossus pictus (Anura).

In Discoglossus pictus eggs, only the dimple contains ionic channels active at fertilization; in particular, chloride channels are found in the central portion of the dimple, which is also the site of sperm penetration. Moreover the dimple hosts an imposing cytoskeleton, consisting of a cortical network and bundles of microfilaments extending from the microvilli. Since spectrin cross links actin and is connected through ankyrin to anion transporters in the plasma membrane of erythrocytes as well as to anion channels in other cells, we studied, in D. pictus egg, the relationship between the localization of spectrin and the high polarization of ionic channels and cytoskeletal organization. By means of immunocytochemistry, we localized spectrin exclusively in the egg dimple. In an attempt to trace back the source of spectrin localization, we immunostained sections of D. pictus ovary and localized spectrin in the nuclei of previtellogenic oocytes, where actin is also present. Antispectrin staining remained until germinal vesicle breakdown. By contrast, a cortical localization was found only when the oocytes divided into two hemispheres and into the germinative area (GA), which, after germinal vesicle breakdown, gives rise to the dimple. At this stage the antispectrin signal was particularly strong in the GA. Using Rho-pialloidin, we also established that spectrin is generally present where F-actin is found. However, spectrin and F-actin do not have the same pattern of fluorescence. In conclusion, our data suggest that spectrin may play a role in oocyte and egg polarity. In eggs, it could be instrumental in anchoring to the cytoskeleton membrane proteins such as receptors and ionic channels, including chloride-permeable channels.

Animals↗